Genetically mapping adipogenesis by using a genome-wide CRISPR-based screen
Abstract/Contents
- Abstract
- Obesity, as characterized by the expansion of white adipose tissue (WAT), has become a global health crisis. The inappropriate storage of excess nutrients in the form of hypertrophic adipocytes is correlated with many metabolic diseases, and their sequelae. Herein, we performed a genome-wide sgRNA CRISPR KO screen in the mouse 3T3-L1 cell line to identify the key genetic regulators with the in vitro proxy for two key features of stem cell biology, the ability to self-renew and differentiate. By using a pooled screen approach followed by FACS to isolate population based on their uptake of a lipophilic dye BODIPY 493/503 we were able to enrich for genes that played key roles during the two phases of 3T3-L1 differentiation: mitotic clonal expansion (MCE) and lipogenesis. These analyses were supplemented by proteomic work, which revealed that changes in protein abundance during differentiation were being largely driven by selective protein translation and degradation, and not transcription. Together, these protein-based and genomic approaches yielded new insights into the specialized translational program driven by the hypusination of Eif5a, and the role of the neddylation/ubiquitin pathway in insulin sensitization within the adipocyte precursor cell. By identifying the genes involved in MCE and lipogenesis, we defined two paths for future therapeutic intervention. We anticipate genes involved in MCE can be used to manipulate the tissue resident preadipocyte to self-renew while genes involved in lipogenesis can be leveraged to control the inappropriate size of the disease correlated hypertrophic adipocytes.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Johnson, Carl Takeshi |
|---|---|
| Degree supervisor | Jackson, Peter K. (Peter Kent) |
| Thesis advisor | Jackson, Peter K. (Peter Kent) |
| Thesis advisor | Majeti, Ravindra, 1972- |
| Thesis advisor | Weissman, Irving L |
| Thesis advisor | Wernig, Marius |
| Degree committee member | Majeti, Ravindra, 1972- |
| Degree committee member | Weissman, Irving L |
| Degree committee member | Wernig, Marius |
| Associated with | Stanford University, Program in Stem Cell Biology and Regenerative Medicine |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Carl Takeshi Johnson. |
|---|---|
| Note | Submitted to the Program in Stem Cell Biology and Regenerative Medicine. |
| Thesis | Thesis Ph.D. Stanford University 2020. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Carl Takeshi Johnson
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